U.S. patent application number 15/494088 was filed with the patent office on 2017-10-26 for multimodality multi-axis 3-d imaging with x-ray.
This patent application is currently assigned to LI-COR, Inc.. The applicant listed for this patent is LI-COR, Inc.. Invention is credited to Han-Wei Wang.
Application Number | 20170309063 15/494088 |
Document ID | / |
Family ID | 58668993 |
Filed Date | 2017-10-26 |
United States Patent
Application |
20170309063 |
Kind Code |
A1 |
Wang; Han-Wei |
October 26, 2017 |
Multimodality Multi-Axis 3-D Imaging With X-Ray
Abstract
Methods and devices are disclosed for the imaging of a
biological sample from all rotational perspectives in
three-dimensional space and with multiple imaging modalities. A
biological sample is positioned on an imaging stage that is capable
of full 360-degree rotation in at least one of two orthogonal axes.
Positioned about the stage are imaging modules enabling the
recording of a series of images in multiple modalities, including
reflected visible light, fluorescence, X-ray, ultrasound, and
optical coherence tomography. A computer can use the images to
construct three-dimensional models of the sample and to render
images of the sample conveying information from one or more imaging
channels. The rendered images can be displayed for an operator who
can manipulate the images to present additional information or
viewing angles of the sample. The image manipulation can be with
touch gestures entered using a sterilizable or disposable touch
pen.
Inventors: |
Wang; Han-Wei; (Lincoln,
NE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
LI-COR, Inc. |
Lincoln |
NE |
US |
|
|
Assignee: |
LI-COR, Inc.
Lincoln
NE
|
Family ID: |
58668993 |
Appl. No.: |
15/494088 |
Filed: |
April 21, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62325588 |
Apr 21, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 21/4795 20130101;
G01N 29/0672 20130101; G01N 2223/408 20130101; G06F 3/04842
20130101; G06F 3/03545 20130101; G01N 29/225 20130101; G01N
2223/401 20130101; G01N 23/223 20130101; G01N 29/2418 20130101;
G06T 19/20 20130101; G06F 3/04886 20130101; G01N 23/2204 20130101;
G06T 19/003 20130101; G06F 3/04845 20130101; G01N 21/6456 20130101;
G01N 23/046 20130101; G01N 21/31 20130101; G06T 15/20 20130101;
G06T 2219/2016 20130101; B43K 8/02 20130101; G01N 21/1702
20130101 |
International
Class: |
G06T 15/20 20110101
G06T015/20; G01N 23/22 20060101 G01N023/22; G01N 23/223 20060101
G01N023/223; G06F 3/0354 20130101 G06F003/0354; G06T 19/00 20110101
G06T019/00; G06F 3/0488 20130101 G06F003/0488; G06F 3/0484 20130101
G06F003/0484; B43K 8/02 20060101 B43K008/02; G06F 3/0484 20130101
G06F003/0484; G06T 19/20 20110101 G06T019/20 |
Claims
1. An apparatus for imaging a biological sample, the apparatus
comprising: a rotatable imaging stage adapted for supporting at
least a portion of a biological sample within an imaging volume,
the rotatable imaging stage comprising a first rotational axis and
a second rotational axis, wherein the second rotational axis is
orthogonal to the first rotational axis; an X-ray source configured
to irradiate the imaging volume with X-rays; an X-ray imager
configured to detect X-rays exiting the imaging volume; a
fluorescence excitation light source configured to illuminate the
imaging volume; a first camera configured to have a depth of focus
within the imaging volume and to detect reflected light; and a
second camera configured to have a depth of focus within the
imaging volume and to detect fluorescence.
2. The apparatus of claim 1, wherein the imaging stage comprises a
transparent portion that is transparent to visible light and
near-infrared light.
3. The apparatus of claim 2, wherein the transparent portion is
transparent to X-rays.
4. The apparatus of claim 2, wherein the transparent portion
comprises glass or acrylic.
5. The apparatus of claim 1, wherein the imaging stage comprises a
plurality of marks at predetermined intervals, wherein the marks
comprise a radiopaque material.
6. The apparatus of claim 5, wherein the radiopaque material
comprises a metal.
7. The apparatus of claim 1, wherein the X-ray source is an X-ray
tube.
8. The apparatus of claim 1, wherein the X-ray imager is a flat
panel detector.
9. The apparatus of claim 1, wherein the first camera is the second
camera.
10. The apparatus of claim 1, further comprising a computer
processer operatively connected with a machine-readable
non-transitory medium embodying information indicative of
instructions for causing the computer processor to perform
operations comprising: recording reflected light images of the
biological sample using the first camera; recording fluorescence
images of the biological sample using the second camera; recording
X-ray images of the biological sample using the X-ray imager; and
rotating the rotatable imaging stage around at least one of the
first rotational axis and the second rotational axis.
11. The apparatus of claim 10, wherein the operations further
comprise: constructing a three-dimensional reflected light model
from two or more reflected light images, wherein each of the two or
more reflected light images is recorded with the rotatable imaging
stage oriented in different positions around at least one of the
first rotational axis and the second rotational axis; constructing
a three-dimensional fluorescence model from two or more
fluorescence images, wherein each of the two or more fluorescence
images is recorded with the rotatable imaging stage oriented in
different positions around at least one of the first rotational
axis and the second rotational axis; construction a
three-dimensional X-ray model from two or more X-ray images,
wherein each of the two or more X-ray images is recorded with the
rotatable imaging stage oriented in different positions around at
least one of the first rotational axis or the second rotational
axis; and rendering an image produced from the reflected light
model, the fluorescence model, and the X-ray model, wherein the
reflected light model, the fluorescence model, and the X-ray model
are identically registered in three-dimensional space.
12. The apparatus of claim 10, wherein the operations further
comprise: associating a first X-ray image of the X-ray images with
a first reflected light image of the reflected light images and a
first fluorescence image of the fluorescence images based on angles
of the first and second rotational axes; rendering a combined image
based on the first X-ray, first reflected light, and first
fluorescence images; and displaying the combined image to a user in
series with other combined images.
13.-33. (canceled)
34. A method for imaging a biological sample, the method
comprising: illuminating a biological sample within an imaging
volume on a rotatable imaging stage with visible light, the
rotatable imaging stage comprising a first rotational axis, a
second rotational axis, and a transparent portion, wherein the
second rotational axis is orthogonal to the first rotational axis,
and wherein the transparent portion is transparent to visible
light, near-infrared light, and X-rays; recording using a first
camera a first reflected light image of visible light reflected by
the biological sample; illuminating the biological sample on the
rotatable imaging stage with fluorescence excitation light using a
fluorescence excitation light source; recording using a second
camera a first fluorescence image of fluorescence emission light
emitted by the biological sample; irradiating the biological sample
on the rotatable imaging stage with X-rays using an X-ray source;
recording using an X-ray imager a first X-ray image of the X-rays
exiting the imaging volume; rotating the imaging stage by a
predetermined amount around at least one of the first rotational
axis and the second rotational axis; recording a second reflected
light image of visible light reflected by the biological sample
through the transparent portion of the rotatable imaging stage;
illuminating the biological sample with fluorescence excitation
light; recording a second fluorescence image of fluorescence
emission light emitted by the biological sample through the
transparent portion of the rotatable imaging stage; irradiating the
biological sample with X-rays; and recording a second X-ray image
of the X-rays exiting the imaging volume through the transparent
portion of the rotatable imaging stage.
35. The method of claim 34 further comprising: constructing a
three-dimensional reflected light model from the first and second
reflected light images using a computer; constructing a
three-dimensional fluorescence model from the first and second
fluorescence images using the computer; constructing a
three-dimensional X-ray model from the first and second X-ray
images using the computer; and rendering an image produced from the
reflected light model, the fluorescence model, and the X-ray model,
wherein the reflected light model, the fluorescence model, and the
X-ray model are identically registered in three-dimensional
space.
36. The method of claim 34 or 35 further comprising: positioning
the X-ray imager between the biological sample and the camera,
wherein the X-ray imager is a flat panel detector, wherein the flat
panel detector has a detection face and a display face, wherein the
display face is opposite to the detection face, wherein the
detection face is directed towards the biological sample, and
wherein the display face is directed towards the camera;
irradiating the biological sample on the rotatable imaging stage
with X-rays using an X-ray source, wherein the biological sample is
positioned between the X-ray source and the flat panel detector,
and wherein the X-ray source, the biological sample, the flat panel
detector, and the first camera are collinear; converting the X-rays
detected by the detection face of the flat panel detector into a
first X-ray image displayed on the display face of the flat panel
detector; recording using the first camera the first X-ray image
displayed on the display face of the flat panel detector;
positioning the flat panel detector such that the flat panel
detector is not between the biological sample and the camera;
rotating the imaging stage by a predetermined amount around at
least of the first rotational axis and the second rotational axis;
positioning the flat panel detector between the biological sample
and the camera; irradiating the biological sample on the rotatable
imaging stage with X-rays using an X-ray source; converting the
X-rays detected through the transparent portion of the rotatable
imaging stage by the detection face of the flat panel detector into
a second X-ray image displayed on the display face of the flat
panel detector; and recording using the first camera the second
X-ray image displayed on the display face of the X-ray flat panel
detector.
37.-44. (canceled)
45. A method of presenting to an operator an image on a
two-dimensional display, the method comprising: displaying on the
two-dimensional display an image, wherein the image is a view of a
subject from a viewpoint, wherein the subject comprises a first
rotational axis and a second rotational axis, wherein the second
rotational axis is orthogonal to the first rotational axis, wherein
the image is produced from two or more three-dimensional models,
wherein the models are each constructed from two or more images of
the subject, wherein at least one of the models is a fluorescence
model constructed from two or more fluorescence images of the
subject, and wherein the models are identically registered in
three-dimensional space; changing the displayed image to a view of
the subject from a viewpoint that is closer to the subject in
response to a zoom command by the operator; changing the displayed
image to a view of the subject from a viewpoint that is farther
from the subject in response to a pinch command by the operator;
changing the displayed image to a view of the subject from a
viewpoint that is rotated around the first rotational axis in
response to a first rotational command by the operator; changing
the displayed image to a view of the subject from a viewpoint that
is rotated around the second rotational axis in response to a
second rotational command by the operator; and displaying
information associated with at least a portion of the displayed
image in response to a selection command by the operator; wherein
at least one of the zoom, pinch, rotational, or selection commands
is entered using touch gestures with a touch pen; wherein the touch
pen comprises a pen body, a touch tip, and an ink tip; wherein the
touch tip is attached to a first end of the pen body; wherein the
ink tip is attached to a second end of the pen body; wherein the
second end is opposite to the first end; wherein the ink tip is
configured to dispense ink; wherein the ink comprises a fluorescent
dye having an emission wavelength; and wherein the fluorescence
images are images of light having the emission wavelength.
46. The method of claim 45, wherein one or more of the zoom, pinch,
rotational, or selection commands are entered using key presses,
control sticks, touch gestures, voice activation, or
accelerometers.
47. The method of claim 45, wherein the touch pen is sterile.
48. The method of claim 45, wherein the pen body comprises
stainless steel.
49. The method of claim 45, wherein the touch tip is detachable
from the pen body and replaceable with a second touch tip.
50.-52. (canceled)
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 62/325,588, filed Apr. 21, 2016, which is
incorporated in its entirety herein for all purposes.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH AND DEVELOPMENT
[0002] NOT APPLICABLE
BACKGROUND
[0003] Assessment of tumor margin during surgery is essential to
the optimal outcome of many oncologic procedures. Tumor margins are
the healthy tissue surrounding the tumor, more specifically, the
distance between the tumor tissue and the edge of the surrounding
tissue removed along with the tumor. Ideally, the margins are
selected so that the risk of leaving tumor tissue within the
patient is low.
[0004] Fluorescence image-guided systems can be used in conjunction
with a series of imaging agents to visualize tumor margins during
surgical procedures for cancer removal. However, in many cancer
surgeries deep surgical cavities with closed spaces and hidden
linings pose significant challenges for the use of over-head
imaging systems. This is particularly true for breast-conserving
surgeries and treatments of head and neck cancers. Discharging
bio-fluids and small fields of view also can compromise the utility
of handheld fluorescence devices for margin assessment at the
surgical cavity. Therefore, intraoperative diagnosis on resected
surgical samples promises to be a more effective means for margin
assessment in many surgical cancer treatment applications.
BRIEF SUMMARY
[0005] In general, provided herein are devices and methods for
imaging with multiple imaging modalities to provide views of the
sample from positions distributed about the sample in any
three-dimensional rotational direction. The sample is positioned on
an imaging stage that is transparent to visible light,
near-infrared light, X-rays, or other types of radiation relevant
to the imaging modalities being used. The imaging stage is fully
rotatable in any direction, so that cameras, detectors, or sensors
located at positions about the stage can record images of the
sample taken from multiple angles. Because the imaging stage is
transparent, these images of the sample can be recorded through the
stage itself.
[0006] Typical imaging modalities used include full-color or
black-and-white imaging of the sample with cameras that record
reflected visible light, and fluorescence imaging with cameras that
record regions of the sample that fluoresce when illuminated at an
excitation frequency. This fluorescence can be associated with dyes
that have binding affinity for diseased cells to be surgically
removed. Other imaging modalities that can be used include X-ray
imaging to visualize tissue density and radiopaque tissue inserts,
photoacoustic imaging, thermoacoustic imaging, ultrasonic imaging,
and optical coherence tomography (OCT).
[0007] A display device can be used to present images rendered
using information recorded in each of the multiple imaging
modalities. The presentation can simultaneously or sequentially
display images from multiple modalities or from multiple angles
relative to the sample. This multimodal and multi-axis imaging can,
for example, offer a novel way to visualize resected tissue
samples, providing surgeons with an improved understanding of tumor
outlines and tissue characteristics.
[0008] One provided apparatus for imaging a biological sample with
visible light, fluorescence, and X-rays, comprises a rotatable
imaging stage adapted for supporting at least a portion of a
biological sample within an imaging volume. The rotatable imaging
stage has a first rotational axis and a second rotational axis. The
second rotational axis is orthogonal to the first rotational axis.
The apparatus further comprises an X-ray source configured to
irradiate the imaging volume with X-rays, an X-ray imager
configured to detect X-rays exiting the imaging volume, a
fluorescence excitation light source configured to illuminate the
imaging volume, a first camera configured to have a depth of focus
within the imaging volume and to detect reflected light, and a
second camera configured to have a depth of focus within the
imaging volume and to detect fluorescence.
[0009] In some embodiments, the imaging stage comprises a
transparent portion that is transparent to visible light and
near-infrared light. In some embodiments, the transparent portion
is transparent to X-rays. In some embodiments, the transparent
portion comprises glass or acrylic. In some embodiments the imaging
stage comprises a plurality of marks at predetermined intervals,
wherein the marks comprise a radiopaque material. In some
embodiments, the radiopaque material comprises a metal.
[0010] In some embodiments, the X-ray source is an X-ray tube. In
some embodiments, the X-ray imager is a flat panel detector. In
some embodiments, the first camera is the second camera.
[0011] In some embodiments, the apparatus further comprises a
computer processer operatively connected with a machine-readable
non-transitory medium embodying information indicative of
instructions for causing the computer processor to perform
operations. In some embodiments, the computer processor records
reflected light images of the biological sample using the first
camera. In some embodiments, the computer processor records
fluorescence images of the biological sample using the second
camera. In some embodiments, the computer processor records X-ray
images of the biological sample using the X-ray imager. In some
embodiments, the computer processor rotates the rotatable imaging
stage around at least one of the first rotational axis and the
second rotational axis.
[0012] In some embodiments, the computer processor constructs a
three-dimensional reflected light model from two or more reflected
light images, wherein each of the two or more reflected light
images is recorded with the rotatable imaging stage oriented in
different positions around at least one of the first rotational
axis and the second rotational axis. In some embodiments, the
computer processor constructs a three-dimensional fluorescence
model from two or more fluorescence images, wherein each of the two
or more fluorescence images is recorded with the rotatable imaging
stage oriented in different positions around at least one of the
first rotational axis and the second rotational axis. In some
embodiments, the computer processor constructs a three-dimensional
X-ray model from two or more X-ray images, wherein each of the two
or more X-ray images is recorded with the rotatable imaging stage
oriented in different positions around at least one of the first
rotational axis or the second rotational axis. In some embodiments,
the computer processor renders an image produced from the reflected
light model, the fluorescence model, and the X-ray model, wherein
the reflected light model, the fluorescence model, and the X-ray
model are identically registered in three-dimensional space.
[0013] Also provided is an apparatus for imaging a biological
sample with visible light, fluorescence, and ultrasound, comprising
a rotatable imaging stage adapted for supporting at least a portion
of a biological sample within an imaging volume. The rotatable
imaging stage comprises a first rotational axis and a second
rotational axis. The second rotational axis is orthogonal to the
first rotational axis. The apparatus further comprises an energy
source configured to transmit energy pulses into the imaging
volume, an ultrasonic transducer array configured to detect
ultrasonic emissions exiting the imaging volume, a fluorescence
excitation light source configured to illuminate the imaging
volume, a first camera configured to have a depth of focus within
the imaging volume and to detect reflected light, and a second
camera configured to have a depth of focus within the imaging
volume and to detect fluorescence.
[0014] In some embodiments, the energy pulses are non-ionizing
laser pulses. In some embodiments, the energy pulses are radio
frequency pulses. In some embodiments, the energy pulses are
ultrasonic pulses.
[0015] In some embodiments, the imaging stage comprises a
transparent portion that is transparent to visible light and
near-infrared light. In some embodiments, the imaging stage
comprises a plurality of marks at predetermined intervals. In some
embodiments, the first camera is the second camera.
[0016] In some embodiments, the apparatus further comprises a
computer processer operatively connected with a machine-readable
non-transitory medium embodying information indicative of
instructions for causing the computer processor to perform
operations. In some embodiments, the computer processor records
reflected light images of the biological sample using the first
camera. In some embodiments, the computer processor records
fluorescence images of the biological sample using the second
camera. In some embodiments, the computer processor records
ultrasound images of the biological sample using the ultrasonic
transducer array. In some embodiments, the computer processor
rotates the rotatable imaging stage around at least one of the
first rotational axis and the second rotational axis.
[0017] In some embodiments, the computer processor constructs a
three-dimensional reflected light model from two or more reflected
light images, wherein each of the two or more reflected light
images is recorded with the rotatable imaging stage oriented in
different positions around at least one of the first rotational
axis and the second rotational axis. In some embodiments, the
computer processor constructs a three-dimensional fluorescence
model from two or more fluorescence images, wherein each of the two
or more fluorescence images is recorded with the rotatable imaging
stage oriented in different positions around at least one of the
first rotational axis and the second rotational axis. In some
embodiments, the computer processor constructs a three-dimensional
ultrasonic model from two or more ultrasound images, wherein each
of the two or more ultrasound images is recorded with the rotatable
imaging stage oriented in different positions around at least one
of the first rotational axis or the second rotational axis. In some
embodiments, the computer processor renders an image produced from
the reflected light model, the fluorescence model, and the
ultrasonic model, wherein the reflected light model, the
fluorescence model, and the ultrasonic model are identically
registered in three-dimensional space.
[0018] Also provided is an apparatus for imaging a biological
sample with visible light, fluorescence, and optical coherence
tomography, comprising a rotatable imaging stage adapted for
supporting at least a portion of a biological sample within an
imaging volume. The rotatable imaging stage has a first rotational
axis and a second rotational axis. The second rotational axis is
orthogonal to the first rotational axis. The apparatus further
comprises a near-infrared light source configured to transmit
near-infrared light into the imaging volume, a fluorescence
excitation light source configured to illuminate the imaging
volume, a first camera configured to have a depth of focus within
the imaging volume and to detect reflected light, a second camera
configured to have a depth of focus within the imaging volume and
to detect fluorescence, and a third camera configured to have a
depth of focus within the imaging volume and to detect
near-infrared light.
[0019] In some embodiments, the imaging stage comprises a
transparent portion that is transparent to visible light and
near-infrared light. In some embodiments, the imaging stage
comprises a plurality of marks at predetermined intervals. In some
embodiments, the first camera is the second camera.
[0020] In some embodiments, the apparatus further comprises a
computer processer operatively connected with a machine-readable
non-transitory medium embodying information indicative of
instructions for causing the computer processor to perform
operations. In some embodiments, the computer processor records
reflected light images of the biological sample using the first
camera. In some embodiments, the computer processor records
fluorescence images of the biological sample using the second
camera. In some embodiments, the computer processor records optical
coherence tomography images of the biological sample using the
third camera. In some embodiments, the computer processor rotates
the rotatable imaging stage around at least one of the first
rotational axis and the second rotational axis.
[0021] In some embodiments, the computer processor constructs a
three-dimensional reflected light model from two or more reflected
light images, wherein each of the two or more reflected light
images is recorded with the rotatable imaging stage oriented in
different positions around at least one of the first rotational
axis and the second rotational axis. In some embodiments, the
computer processor constructs a three-dimensional fluorescence
model from two or more fluorescence images, wherein each of the two
or more fluorescence images is recorded with the rotatable imaging
stage oriented in different positions around at least one of the
first rotational axis and the second rotational axis. In some
embodiments, the computer processor constructs a three-dimensional
optical coherence tomography model from two or more optical
coherence tomography images, wherein each of the two or more
optical coherence tomography images is recorded with the rotatable
imaging stage oriented in different positions around at least one
of the first rotational axis or the second rotational axis. In some
embodiments, the computer processor renders an image produced from
the reflected light model, the fluorescence model, and the optical
coherence tomography model, wherein the reflected light model, the
fluorescence model, and the optical coherence tomography model are
identically registered in three-dimensional space.
[0022] Also provided is a sterile touch pen comprising a pen body
and a pen tip. The pen tip is attached to an end of the pen body.
In some embodiments, the pen tip is a touch tip, the end of the pen
body is a first end, and the touch pen further comprises an ink tip
configured to dispense ink, wherein the ink tip is attached to a
second end of the pen body, and wherein the second end is opposite
to the first end. In some embodiments, the ink comprises a
fluorescent dye. In some embodiments, the pen body comprises
stainless steel. In some embodiments, the pen tip is detachable
from the pen body and replaceable with a second pen tip. In some
embodiments, the touch pen further comprises a pen cover. The pen
cover encloses the pen body and the pen tip, and is sterile.
[0023] Also provided is a method for imaging a biological sample
with visible light, fluorescence and X-rays. The method comprises
illuminating a biological sample within an imaging volume on a
rotatable imaging stage with visible light. The rotatable imaging
stage has a first rotational axis, a second rotational axis, and a
transparent portion. The second rotational axis is orthogonal to
the first rotational axis. The transparent portion is transparent
to visible light, near-infrared light, and X-rays. The method
further comprises recording, using a first camera, a first
reflected light image of visible light reflected by the biological
sample. The method further comprises illuminating the biological
sample on the rotatable imaging stage with fluorescence excitation
light using a fluorescence excitation light source. The method
further comprises recording, using a second camera, a first
fluorescence image of fluorescence emission light emitted by the
biological sample. The method further comprises irradiating the
biological sample on the rotatable imaging stage with X-rays using
an X-ray source. The method further comprises recording, using an
X-ray imager, a first X-ray image of the X-rays exiting the imaging
volume. The method further comprises rotating the imaging stage by
a predetermined amount around at least one of the first rotational
axis and the second rotational axis. The method further comprises
recording a second reflected light image of visible light reflected
by the biological sample through the transparent portion of the
rotatable imaging stage. The method further comprises illuminating
the biological sample with fluorescence excitation light. The
method further comprises recording a second fluorescence image of
fluorescence emission light emitted by the biological sample
through the transparent portion of the rotatable imaging stage. The
method further comprises irradiating the biological sample with
X-rays. The method further comprises recording a second X-ray image
of the X-rays exiting the imaging volume through the transparent
portion of the rotatable imaging stage.
[0024] In some embodiments, the method further comprises
constructing a three-dimensional reflected light model from the
first and second reflected light images using a computer. In some
embodiments, the method further comprises constructing a
three-dimensional fluorescence model from the first and second
fluorescence images using the computer. In some embodiments, the
method further comprises constructing a three-dimensional X-ray
model from the first and second X-ray images using the computer. In
some embodiments, the method further comprises rendering an image
produced from the reflected light model, the fluorescence model,
and the X-ray model, wherein the reflected light model, the
fluorescence model, and the X-ray model are identically registered
in three-dimensional space.
[0025] In some embodiments, the method further comprises
positioning the X-ray imager between the biological sample and the
camera. In some embodiments, the X-ray imager is a flat panel
detector. In some embodiments, the flat panel detector has a
detection face and a display face, wherein the display face is
opposite to the detection face, wherein the detection face is
directed towards the biological sample, and wherein the display
face is directed towards the camera. In some embodiments, the
method further comprises irradiating the biological sample on the
rotatable imaging stage with X-rays using an X-ray source, wherein
the biological sample is positioned between the X-ray source and
the flat panel detector, and wherein the X-ray source, the
biological sample, the flat panel detector, and the first camera
are collinear. In some embodiments, the method further comprises
converting the X-rays detected by the detection face of the flat
panel detector into a first X-ray image displayed on the display
face of the flat panel detector. In some embodiments, the method
further comprises recording using the first camera the first X-ray
image displayed on the display face of the flat panel detector. In
some embodiments, the method further comprises positioning the flat
panel detector such that the flat panel detector is not between the
biological sample and the camera. In some embodiments, the method
further comprises rotating the imaging stage by a predetermined
amount around at least of the first rotational axis and the second
rotational axis. In some embodiments, the method further comprises
positioning the flat panel detector between the biological sample
and the camera. In some embodiments, the method further comprises
irradiating the biological sample on the rotatable imaging stage
with X-rays using an X-ray source. In some embodiments, the method
further comprises converting the X-rays detected through the
transparent portion of the rotatable imaging stage by the detection
face of the flat panel detector into a second X-ray image displayed
on the display face of the flat panel detector. In some
embodiments, the method further comprises recording, using the
first camera, the second X-ray image displayed on the display face
of the X-ray flat panel detector.
[0026] Also provided is a method for imaging a biological sample
with visible light, fluorescence, and ultrasound. The method
comprises illuminating a biological sample within an imaging volume
on a rotatable imaging stage with visible light. The rotatable
imaging stage has a first rotational axis, a second rotational
axis, and a transparent portion. The second rotational axis is
orthogonal to the first rotational axis. The transparent portion is
transparent to visible light and near-infrared light. The method
further comprises recording using a first camera a first reflected
light image of visible light reflected by the biological sample.
The method further comprises illuminating the biological sample on
the rotatable imaging stage with fluorescence excitation light
using a fluorescence excitation light source. The method further
comprises recording using a second camera a first fluorescence
image of fluorescence emission light emitted by the biological
sample. The method further comprises transmitting energy pulses
into the biological sample, wherein the energy pulses are absorbed
by the biological sample and converted into ultrasonic emissions.
The method further comprises detecting the ultrasonic emissions
using an ultrasonic transducer array. The method further comprises
recording a first ultrasound image constructed from the ultrasonic
emissions detected by the ultrasonic transducer array. The method
further comprises rotating the imaging stage by a predetermined
amount around at least one of the first rotational axis and the
second rotational axis. The method further comprises recording a
second reflected light image of visible light reflected by the
biological sample through the transparent portion of the rotatable
imaging stage. The method further comprises illuminating the
biological sample with fluorescence excitation light. The method
further comprises recording a second fluorescence image of
fluorescence emission light emitted by the biological sample
through the transparent portion of the rotatable imaging stage. The
method further comprises transmitting energy pulses into the
biological sample, wherein the energy pulses are absorbed by the
biological sample and converted into ultrasonic emissions. The
method further comprises detecting the ultrasonic emissions using
an ultrasonic transducer array. The method further comprises
recording a second ultrasound image constructed from the ultrasonic
emissions detected by the ultrasonic transducer array.
[0027] In some embodiments, the method further comprises
constructing a three-dimensional reflected light model from the
first and second reflected light images using a computer. In some
embodiments, the method further comprises constructing a
three-dimensional fluorescence model from the first and second
fluorescence images using the computer. In some embodiments, the
method further comprises constructing a three-dimensional
ultrasonic model from the first and second ultrasound images using
the computer. In some embodiments, the method further comprises
rendering an image produced by overlaying the reflected light
model, the fluorescence model, and the ultrasonic model, wherein
the reflected light model, the fluorescence model, and the
ultrasonic model are identically registered in three-dimensional
space.
[0028] In some embodiments, the energy pulses are non-ionizing
laser pulses and the ultrasound image is a photoacoustic image. In
some embodiments, the energy pulses are radio frequency pulses and
the ultrasound image is a thermoacoustic image. In some
embodiments, the energy pulses are ultrasonic pulses.
[0029] Also provided is a method for imaging a biological sample
with visible light, fluorescence, and optical coherence tomography.
The method comprises illuminating a biological sample within an
imaging volume on a rotatable imaging stage with visible light. The
rotatable imaging stage has a first rotational axis, a second
rotational axis, and a transparent portion. The second rotational
axis is orthogonal to the first rotational axis. The transparent
portion is transparent to visible light and near-infrared light.
The method further comprises recording using a first camera a first
reflected light image of visible light reflected by the biological
sample. The method further comprises illuminating the biological
sample on the rotatable imaging stage with fluorescence excitation
light using a fluorescence excitation light source. The method
further comprises recording using a second camera a first
fluorescence image of fluorescence emission light emitted by the
biological sample. The method further comprises illuminating the
biological sample on the rotatable imaging stage with near-infrared
light. The method further comprises recording using a third camera
a first optical coherence tomography image of near-infrared light
reflected by the biological sample. The method further comprises
rotating the imaging stage by a predetermined amount around at
least one of the first rotational axis and the second rotational
axis. The method further comprises recording a second reflected
light image of visible light reflected by the biological sample
through the transparent portion of the rotatable imaging stage. The
method further comprises illuminating the biological sample with
fluorescence excitation light. The method further comprises
recording a second fluorescence image of fluorescence emission
light emitted by the biological sample through the transparent
imaging stage. The method further comprises illuminating the
biological sample with near-infrared light. The method further
comprises recording a second optical coherence tomography image of
near-infrared light reflected by the biological sample
[0030] In some embodiments, the method further comprises
constructing a three-dimensional reflected light model from the
first and second reflected light images using a computer. In some
embodiments, the method further comprises constructing a
three-dimensional fluorescence model from the first and second
fluorescence images using the computer. In some embodiments, the
method further comprises constructing a three-dimensional optical
coherence tomography model from the first and second optical
coherence tomography images using the computer. In some
embodiments, the method further comprises rendering an image
produced from the reflected light model, the fluorescence model,
and the optical coherence tomography model, wherein the reflected
light model, the fluorescence model, and the optical coherence
tomography model are identically registered in three-dimensional
space.
[0031] In some embodiments, the second camera is the first
camera.
[0032] Also provided is a method of presenting to an operator an
image on a two-dimensional display. The method comprises displaying
on the two-dimensional display an image. The image is a view of a
subject from a viewpoint. The subject has a first rotational axis
and a second rotational axis. The second rotational axis is
orthogonal to the first rotational axis. The method further
comprises changing the displayed image to a view of the subject
from a viewpoint that is closer to the subject in response to a
zoom command by the operator. The method further comprises changing
the displayed image to a view of the subject from a viewpoint that
is farther from the subject in response to a pinch command by the
operator. The method further comprises changing the displayed image
to a view of the subject from a viewpoint that is rotated around
the first rotational axis in response to a first rotational command
by the operator. The method further comprises changing the
displayed image to a view of the subject from a viewpoint that is
rotated around the second rotational axis in response to a second
rotational command by the operator. The method further comprises
displaying information associated with at least a portion of the
displayed image in response to a selection command by the
operator.
[0033] In some embodiments, the image is produced from two or more
three-dimensional models. In some embodiments, the models are each
constructed from two or more images of the subject, and are each
identically registered in three-dimensional space.
[0034] In some embodiments one or more of the zoom, pinch,
rotational, or selection commands are entered using key presses,
control sticks, touch gestures, voice activation, or
accelerometers.
[0035] In some embodiments, the touch gestures are entered using a
touch pen. The touch pen comprises a pen body and a pen tip. The
pen tip is attached to an end of the pen body. In some embodiments,
the pen tip is a touch tip, the end of the pen body is a first end,
and the touch pen further comprises an ink tip configured to
dispense ink, wherein the ink tip is attached to a second end of
the pen body, and wherein the second end is opposite to the first
end. In some embodiments, the ink comprises a fluorescent dye. In
some embodiments, the touch pen is sterile. In some embodiments,
the pen body comprises stainless steel. In some embodiments, the
pen tip is detachable from the pen body and replaceable with a
second pen tip. In some embodiments, the touch pen further
comprises a pen cover, wherein the pen cover encloses the pen body
and the pen tip, and wherein the pen cover is sterile.
BRIEF DESCRIPTION OF THE DRAWINGS
[0036] FIG. 1 is a perspective illustration of a rotatable stage
and X-ray imaging system in accordance with an embodiment.
[0037] FIG. 2 is a flowchart of a process in accordance with an
embodiment.
[0038] FIG. 3 is a perspective illustration of an imaging system
with X-ray flat panel detector in accordance with an
embodiment.
[0039] FIG. 4 is a flowchart of a process in accordance with an
embodiment.
[0040] FIG. 5 is a flowchart of a process in accordance with an
embodiment.
[0041] FIG. 6 is a flowchart of a process in accordance with an
embodiment.
[0042] FIG. 7 is an illustration of a touch pen in accordance with
an embodiment.
DETAILED DESCRIPTION
[0043] The present invention relates in part to multimodal and
multi-axis three-dimensional imaging devices and methods for
visualizing samples. The devices and methods can be used to record
and display multimodal images of a biological sample representing
views of the sample from any position rotated about the sample in
three-dimensional space.
[0044] A technical advantage of the embodiments described herein is
that a surgeon can have enhanced access to visualized information
regarding the location and characteristics of diseased and healthy
cells and tissue within a resected biopsy sample. This can allow a
surgeon to more accurately assess tumor margins during surgical
procedures, which can in turn increase the probability of a
successful surgery and the survival rate of the patient.
[0045] By combining multi-axis rotation three-dimensional imaging
with multiple imaging modalities, the inventors have made the
surprising discovery of a novel way to look at a resected tissue
sample. For example, by combining reflected visible light imaging
and fluorescence imaging with an X-ray imaging module, one
embodiment of the system describe herein provides three-dimensional
full-rotation surface mapping together with X-ray projections from
different angles. This can give a surgeon a unique and
comprehensive understanding of the molecular signal from a tumor
tissue from the optical channels together with tomographic
information from the X-ray channel. The optical channels give the
molecular signal of a tumor and the outline of the tissue, and the
X-ray channel shows the tissue density and projection, as well as
information about any metal or wire inserts placed inside the
tissue. In some cases, with a molecular probe that gives contrast
in multiple channels, such as the fluorescence and X-ray channels,
an overlay image of the multimodalities can show signals from the
same imaging agent.
[0046] FIG. 1 illustrates one embodiment as a descriptive example.
Shown is an apparatus 100 comprising a sample positioning module
101 and an optical imaging module 102. The sample positioning
module has an imaging stage 103 that has a first rotational axis
104 and a second rotational axis 105. A biological sample 106 is
shown being supported by the imaging stage 103. The biological
sample 106 is within an imaging volume 107 proximate to the imaging
stage 103. An X-ray source 108 is configured to irradiate the
imaging volume 107 with X-rays, and an X-ray imager 109 is
configured to detect X-rays exiting the imaging volume 107. The
optical imaging module 102 has a fluorescence excitation light
source 110 configured to illuminate the imaging volume 107, and a
camera 111 configured to have a depth of focus within the imaging
volume.
[0047] The biological sample can comprise material removed from a
subject. The subject is typically a human, but also can be another
animal. The subject can be, for example, rodent, canine, feline,
equine, ovine, porcine, or another primate. The subject can be a
patient suffering from a disease. In some embodiments, the subject
is a cancer patient. In certain aspects, the biological sample
comprises a tumor, such as tumor tissue or cells. In certain
aspects, the biological sample comprises a peripheral biopsy of a
tissue sample previously removed. In another aspect, the biological
sample is tumor tissue such as a breast core biopsy. The biological
sample size can be as small as a tissue slice.
[0048] The rotatable imaging stage supporting the biological sample
is equipped with rotational motors and stages to control the view
angle and position of a sample within the imaging volume. By
rotating a sample in two degrees of freedom, the stage can allow an
imager to efficiently provide a full-rotation three-dimensional
image. A first rotational axis can, for example, provide 360-degree
movement along the z-axis (roll) relative to the sample. A second
rotational axis can, for example, move along the y-axis (tilt) for
imaging at different perspectives. Tilting of the sample stage also
allows projection views from the top and the bottom of the sample
via a transparent window. In some embodiments, the rotational
imaging stage can also be moved in an X-Y plane to allow for the
registration of the sample to the center of the imaging volume.
[0049] Rotational combinations can allow the entire sample to be
imaged. To collect pertinent imaging projections along a sample for
subsequent three-dimensional reconstruction, the rotational imaging
stage can rotate the object in rolling and tilting degrees of
freedom. In some embodiments, to provide comprehensive coverage of
sample features the rolling angle is in the range of from 7.5
degrees to 30 degrees, depending on the complexity of the sample.
In some embodiments, a rolling angle of 22.5 degrees and a tilting
angle of .+-.35 degrees offers a full rotation for
three-dimensional inspection and imaging of the sample.
[0050] Rotation of the imaging stage around one or both of the
first and second rotational axis can be accomplished through the
use of rotor bearings connected to the stage. A rotor bearing can
be a hub, axle, or other mechanical element that bears contact
between at least two parts and that allows for rotation around an
axis. A rotary bearing can include circular tracks and cages for
ball bearings, lubricant surfaces, and other friction-reducing
implements.
[0051] The imaging volume is defined as the volume formed by the
fields of illumination or other electromagnetic radiation, by the
depth-of-focus of an object lens, and by the field-of-view of an
imaging head. The imaging volume is typically configured such that
all cameras, detectors, sensors, and other image capturing elements
of the apparatus are tolerant of placement of the sample anywhere
within the volume.
[0052] The X-ray source can be any artificial X-ray source
configured to irradiate the imaging volume with X-rays. In some
embodiments, the X-ray source is an X-ray tube. The X-ray tube can
comprise a rotating anode tube. In some embodiments, the X-ray
source is a solid-anode microfocus X-ray tube or a metal-jet-anode
microfocus X-ray tube.
[0053] The X-ray imager can be any device configured to measure the
properties of X-rays exiting the image volume. The X-ray imager can
comprise, for example, one or more of a sensitized photographic
plate, sensitized photographic film, a photostimulable phosphor
plate, a semiconductor or solid state detector, or a scintillator.
In some embodiments, the X-ray imager comprises a scintillator. The
scintillator can comprise any material that converts an X-ray
photon to a visible light photon. The scintillator can comprise one
or more organic or inorganic compounds. The scintillator compounds
can comprise, for example, barium fluoride, calcium fluoride doped
with europium, bismuth germinate, cadmium tungstate, cesium iodide
doped with thallium, cesium iodide doped with sodium, undoped
cesium iodide, gadolinium oxysulfide, lanthanum bromide doped with
cerium, lanthanum chloride doped with cerium, lead tungstate,
lutetium iodide, lutetium oxyorthosilicate, sodium iodide doped
with thallium, yttrium aluminum garnet, zinc sulfide, or zinc
tungstate. In some embodiments, the scintillator comprises sodium
iodide, gadolinium oxysulfide, or cesium iodide.
[0054] In some embodiments, the X-ray imager is a an X-ray flat
panel detector. The flat panel detector can comprise a scintillator
material and a photodiode transistor array. The flat panel detector
can further comprise one or more readout circuits. The flat panel
detector can comprise a detection face and a display face on
opposite sides of the detector from one another. The detection face
can be directed towards the biological sample and the X-ray source
so as to be contacted with X-rays generated by the X-ray source and
passing through the imaging volume. The display face can be
directed towards a camera so that an X-ray image displayed on the
display face can be recorded using the camera. In some embodiments,
the X-ray image is displayed on the display face by generating
visible light that is recorded by a visible light camera configured
to have a depth of focus that corresponds to the distance between
the display face and the camera.
[0055] In preferred embodiments, the X-ray source, biological
sample, and X-ray imager are collinear with one another. In this
configuration, the X-ray imager can record information related to
X-rays that are generated by the X-ray source, travel through the
imaging volume, and contact the sensors of the X-ray imager. As the
X-rays travel through the imaging volume, they can be affected by
the properties of any material, such as a biological sample, within
the imaging volume. Regions of the biological sample with differing
degrees of radiodensity will permit differing amounts of X-rays to
pass through those regions. These differing amounts will result in
changes in the signal intensities detected by different areas of
the X-ray imager sensors. As the rotatable imaging stage is moved
around one or both of its orthogonal rotational axes, the locations
of any radiodense regions of the biological sample relative to the
locations of the X-ray source and X-ray imager will be changed.
This allows for the recording of X-ray images with the X-ray imager
that provide information about the radiopacity of the sample as
detected from multiple perspectives.
[0056] The fluorescence excitation light source can be any device
configured to emit electromagnetic radiation at an excitation
wavelength capable of exciting a fluorescent material within the
imaging volume. The fluorescent material can comprise a fluorophore
or fluorescent dye. The fluorescence excitation light source is
configured to illuminate the imaging volume, and any sample within,
with radiation comprising this excitation wavelength. In some
embodiments, the fluorescence excitation light source emits
near-infrared light. In certain aspects, the illumination of the
biological sample with near-infrared light is performed at one or
more wavelengths of from about 650 nm to about 1400 nm. These
wavelengths include, for example, about 700, 725, 750, 775, 800,
825, 850, 875, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990,
1000, 1100, 1200, 1300, and 1400 nm. Sometimes these wavelengths
are referred to as being in the NIR-I (between 750 and 1060 nm) and
NIR-II (between 1000 nm and 1700 nm) wavelength regions.
[0057] Fluorophore methods utilize molecules that absorb light of
one wavelength and emit light of a different wavelength. To utilize
a visible image in combination with a fluorophore (e.g., an
infrared or near-infrared fluorophore), care should be taken to
ensure that the spectra of light variously absorbed, reflected, and
emitted do not significantly overlap so as to confound
differentiation of the components from each other and
differentiation of the components from endogenous tissue material.
Filter sets can be used in the optical system to isolate excitation
wavelengths with optimized emission collection for corresponding
imaging agents.
[0058] In certain aspects, the biological sample comprises a
fluorescent dye. In one aspect, the fluorescent group is a
near-infrared (NIR) fluorophore that emits in the range of between
about 650 to about 1400 nm. Use of near-infrared fluorescence
technology is advantageous in the methods herein as it
substantially eliminates or reduces background from auto
fluorescence of tissue. Another benefit to the near-IR fluorescent
technology is that the scattered light from the excitation source
is greatly reduced since the scattering intensity is proportional
to the inverse fourth power of the wavelength. Low background
fluorescence and low scattering result in a high signal to noise
ratio, which is essential for highly sensitive detection.
Furthermore, the optically transparent window in the near-IR region
(650 nm to 990 nm) or NIR-II region (between about 1000 nm and
1400) in biological tissue makes NIR fluorescence a valuable
technology for imaging and subcellular detection applications that
require the transmission of light through biological
components.
[0059] In certain aspects, the fluorescent group is preferably
selected form the group consisting of IRDYE.RTM.800RS, IRDYE.RTM.
800CW, IRDYE.RTM. 800, ALEXA FLUOR.RTM. 660, ALEXA FLUOR.RTM. 680,
ALEXA FLUOR.RTM. 700, ALEXA FLUOR.RTM. 750, ALEXA FLUOR.RTM. 790,
Cy5, Cy5.5, Cy7, DY 676, DY680, DY682, and DY780 molecular marker.
In certain aspects, the near infrared group is IRDYE.RTM. 800CW,
IRDYE.RTM. 800, IRDYE.RTM. 700DX, IRDYE.RTM. 700, or Dynomic DY676
molecular marker.
[0060] In certain aspects, the fluorescent dye is contacted with
the biological sample prior to excising the biological sample from
the subject. For example, the dye can be injected or administered
to the subject prior to surgery or after surgery. In certain
aspects, the dye is conjugated to an antibody, ligand, or targeting
moiety or molecule having an affinity to a tumor or recognizes a
tumor antigen. In certain aspects, the fluorescent dye comprises a
targeting moiety. In one aspect, the surgeon "paints" the tumor
with the dye. In certain aspects, the fluorescent dye is contacted
with the biological sample after excising the biological sample
from the subject. In this manner, dye can be contacted to the
tissue at the margins.
[0061] In some aspects, the targeting molecule or moiety is an
antibody that binds an antigen such as a lung cancer cell surface
antigen, a brain tumor cell surface antigen, a glioma cell surface
antigen, a breast cancer cell surface antigen, an esophageal cancer
cell surface antigen, a common epithelial cancer cell surface
antigen, a common sarcoma cell surface antigen, or an osteosarcoma
cell surface antigen.
[0062] Illumination sources can be mounted proximate to the imaging
volume in order to illuminate the sample with white light,
monochrome light, near-infrared light, fluorescence light, or other
electromagnetic radiation. One or more white lights can be used to
illuminate the imaging volume. In some embodiments, the
illumination of the biological sample with visible light is
performed at one or more wavelengths of about 380 nm to about 700
nm. These wavelengths include, for example, about 380, 390, 400,
425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, or about 700
nm. These wavelengths can occur in combination, such as in
broadband white light.
[0063] One or more cameras of the apparatus can have an actively or
passively cooled heat exchanger to maintain imaging sensors at low
temperatures. The imaging sensors can be charge coupled device
imaging sensors. The cooling can prevent optical background noise
such as darkness or blooming. Other approaches for improving camera
sensitivity to compensate for low light levels of fluorescence can
include imaging with a monochrome sensor, long exposure durations,
and electronic noise suppression methods. Exemplary camera and
optical components are described in U.S. Pat. Nos. 7,286,232,
8,220,415, and 8,851,017.
[0064] The rotatable imaging stage can comprise a transparent
portion, such as a window. The window can be transparent at the
working wavelengths for both reflective light and fluorescence
imaging. The transparent portion can further be transparent to
X-rays. To accommodate a large size sample, the window can be
configured to a shape that is wider than either the projection size
of the imaging volume or the footprint of the target sample. A
circle on the window can be used to mark the border of a suggested
imaging area.
[0065] The material of the transparent portion can be, for example,
borosilicate-based glass, acrylic, or other transparent material.
The surface could be treated or coated for optical or surface
functional requirements. Non-limiting examples of these treatments
include those providing anti-reflection, transparency, absorption,
hydrophobic, or hydrophilic properties to the surface.
[0066] The rotatable imaging stage can further comprise one or more
marks. The marks can be regularly spaced or irregularly spaced. The
marks can be configured to provide reference scales to users of the
apparatus. The marks can also provide references to a computer
processor used to analyze and manipulate images recorded of the
sample within the imaging volume. In some embodiments, the marks
comprise a radiopaque material. The radiopaque material can
comprise a polymer or a metal.
[0067] The devices and methods can utilize a computing apparatus
that is programmed or otherwise configured to automate and/or
regulate one or more steps of the methods or features of the
devices provided herein. Some embodiments provide machine
executable code in a non-transitory storage medium that, when
executed by a computing apparatus, implements any of the methods or
operates any of the devices described herein. In some embodiments,
the computing apparatus operates the power source and/or pump
control.
[0068] In some embodiments, the apparatus comprises a computer
processor that can record images of the biological sample. The
recorded images can be reflected light images captured by a camera
configured to detect reflected light. In some embodiments, the
reflected light is visible light. The recorded images can be
fluorescence images captured by a camera configured to detect
fluorescence emission light. In some embodiments, the same camera
is configured to detect both reflected light and fluorescence
emission light. The recorded images can be X-ray images captured by
an X-ray imager. The X-ray images can be captured by a camera
configured to detect light images presented on a display face of an
X-ray flat panel detector. The computer processor can tag the
recorded images with information related to the relative positions
of one or more of cameras, imagers, detectors, or sensors, with
respect to the rotatable imaging stage. The computer process can
tag the recorded images with information related to the rotational
position of the biological sample around either or both of a first
and second orthogonal rotational axes. The locational and
positional tags can use information determined by detecting the
locations and orientations of one or more marks on the rotational
imaging stage.
[0069] In some embodiments, the computer processor can control the
rotation of the rotatable imaging stage. The rotation can be about
one or both of the first and second orthogonal rotational axes. The
rotation can occur simultaneously along with image recording. The
rotation can be stopped during image recording. In some
embodiments, the rotation is from one predetermined position to
another. In some embodiments, the rotation is to a series of
multiple different predetermined positions. The computer can record
images captured in one or more channels or modalities at each
position. As a non-limiting example, the computer can capture a
reflected light image, a fluorescence image, and an X-ray image at
each position that the rotatable imaging stage is moved to. The
computer processor can rotate the imaging stage so that a
transparent portion of the imaging stage is between the sample and
one or more cameras, imagers, detectors, or sensors. Images or
other information can then be recorded of the sample through the
transparent portion of the imaging stage.
[0070] In some embodiments, the computer processer can construct
models based on the recorded images. The models can be
three-dimensional models. The models can comprise series of
discrete images, each recorded as the rotatable imaging stage was
at a different orientation relative to the apparatus element used
in recording the images. The models can further comprise images
constructed by interpolating information contained in discrete
images. In some embodiments, the models are wireframe models
created by translating two or more images into a polygonal mesh.
The models can comprise surface information about the biological
subject. The models can comprise tomographic information about the
biological subject.
[0071] In some embodiments, the computer processer can render
images produced from the constructed models. The rendered images
can be identical to images recorded using the cameras, imagers,
detectors, or sensors. The rendered images can be constructions
based on information in the recorded images. The rendered images
can contain images or information collected with one channel or
modality. The rendered images can overlay images or information
collected with two or more channels or modalities. As a
non-limiting example, a rendered image can overlay reflected light
information showing a visible light view of the biological sample,
fluorescence information showing locations of fluorescing regions
within the biological sample, and X-ray information showing
locations of radiodense regions within the biological sample.
Typically, when a rendered image overlays images or information
from multiple channels, modalities, or models, the models are
identically registered in three-dimensional space so that the image
presents information for each modality as seen from a single
viewpoint.
[0072] The apparatus can further comprise another energy source
configured to deliver energy pulses into the imaging volume. In
some embodiments, the energy source is a laser. In some
embodiments, the energy source is a radio frequency transmitter. In
some embodiments, the energy source is an ultrasound generator. In
some embodiments, the energy pulses are non-ionizing laser pulses.
In some embodiments, the energy pulses are radio frequency pulses.
In some embodiments, the energy pulses are ultrasonic pulses.
[0073] The apparatus can further comprise an ultrasonic transducer
array configured to detect ultrasound waves exiting the imaging
volume and convert the waves into electrical signals. The energy
pulses transmitted into the imaging volume can cause a biological
sample within to absorb this time-varying energy, inducing the
generation of acoustic waves that can be detected by the ultrasonic
transducer array. Within the imaging volume, the ultrasonic
transducer array is in contact with the biological sample via a
coupling medium. The coupling medium can comprise water or gel to
relay ultrasound waves. In some embodiments, the energy pulses are
non-ionizing laser pulses and the ultrasonic transducer array can
be used to record a photoacoustic image. In some embodiments, the
energy pulses are radio frequency pulses and the ultrasonic
transducer array can be used to record a thermoacoustic image. In
some embodiments, the energy pulses are ultrasonic pulses, and the
ultrasonic transducer array can be used to record an ultrasound
image.
[0074] The apparatus can further comprise an interferometer
configured for optical coherence tomography of the biological
sample within the imaging volume. In some embodiments, the
interferometer is a Michelson interferometer. The apparatus can
further comprise a camera configured to detect electromagnetic
radiation emitted from the imaging volume for optical coherence
tomography of the biological sample.
[0075] FIG. 2 presents a flowchart of a process 200 for imaging a
biological sample with reflected visible light, fluorescence, and
X-rays. In operation 201, a biological sample within an imaging
volume on a rotatable imaging stage is illuminated with visible
light, the rotatable imaging stage comprising a first rotational
axis, a second rotational axis, and a transparent portion, wherein
the second rotational axis is orthogonal to the first rotational
axis, and wherein the transparent portion is transparent to visible
light, near-infrared light, and x-rays. In operation 202, a first
reflected light image of visible light reflected by the biological
sample is recorded using a first camera. In operation 203, the
biological sample on the rotatable imaging stage is illuminated
with fluorescence excitation light using a fluorescence excitation
light source. In operation 204, a first fluorescence image of
fluorescence emission light emitted by the biological sample is
recorded using a second camera. In operation 205, the biological
sample on the rotatable imaging stage is irradiated with X-rays
using an X-ray source. In operation 206, a first X-ray image of the
X-rays exiting the imaging volume is recorded using an X-ray
imager. In operation 207, the imaging stage is rotated by a
predetermined amount around at least one of the first rotational
axis and the second rotational axis. In operation 208, a second
reflected light image of visible light reflected by the biological
sample through the transparent portion of the rotatable imaging
stage is recorded. In operation 209, the biological sample is
illuminated with fluorescence excitation light. In operation 210, a
second fluorescence image of fluorescence emission light emitted by
the biological sample through the transparent portion of the
rotatable imaging stage is recorded. In operation 211, the
biological sample is irradiated with X-rays. In operation 212, a
second X-ray image of the X-rays exiting the imaging volume through
the transparent portion of the rotatable imaging stage is
recorded.
[0076] In some embodiments, the method further comprises an
operation to construct a three-dimensional reflected light model
from the first and second reflected light images using a computer.
In some embodiments, the method further comprises an operation to
construct a three-dimensional fluorescence model from the first and
second fluorescence images using the computer. In some embodiments,
the method further comprises an operation to construct a
three-dimensional X-ray model from the first and second X-ray
images using a computer. In some embodiments, the method further
comprises an operation to render an image produced from the
reflected light model, the fluorescence model, and the X-ray model,
wherein the reflected light model, the fluorescence model, and the
X-ray model are identically registered in three-dimensional
space.
[0077] The process presented in FIG. 2 can be carried out with an
apparatus similar or identical to the one presented in FIG. 1. In
some embodiments, the X-ray source 108 and the X-ray imager 109 are
placed orthogonal to the optical imaging module 102. The center of
the field-of-view of the X-ray imager 109 is on the same x-y plane
as the field-of-view of the optical imaging module 102. In some
embodiments, the X-ray source 108 and the X-ray imager 109 are
placed at a defined angle along the z-axis of the imaging volume
while maintaining the center of the field-of-view of the X-ray
imager on the same x-y plane as the field-of-view of the optical
imaging module 109. Therefore, if the optical module 102 is imaging
the biological sample 106 at an angle of 0 degrees around the
rotational axis of the imaging volume, the X-ray module can be used
to record X-ray projection images of the biological sample at an
angle of 90 degrees, 270 degrees, or any other defined angle around
the rotational axis of the imaging volume. After the rotatable
imaging stage has been rotated to a new orientation, and images of
the biological sample have been recorded at this new orientation,
registration of the images recorded with two or more modalities can
be performed to provide co-localized and co-registered image
information.
[0078] The process presented in FIG. 2 can be carried out with an
apparatus in which the X-ray source 108 can irradiate the imaging
volume with X-rays from the same direction as the direction of the
optical module 102. In this case, the X-ray projection image
recorded using the X-ray imager 109 is a view of the biological
sample 106 from approximately the same angle as that of the image
recorded using the optical module 102. In this embodiment, surface
mapping images recorded using the optical module 102 will have
registrations approximately identical to tomography images
simultaneously recorded using the X-ray module. As a result, an
operation involving subsequent co-localization and co-registration
of the images recorded using different modalities can be
eliminated.
[0079] In some embodiments, the method further comprises an
operation to position the X-ray imager between the biological
sample and the camera, wherein the X-ray imager is a flat panel
detector, wherein the flat panel detector has a detection face and
a display face, wherein the display face is opposite to the
detection face, wherein the detection face is directed towards the
biological sample, and wherein the display face is directed towards
the camera. In some embodiments, the method further comprises an
operation to irradiate the biological sample on the rotatable
imaging stage with X-rays using an X-ray source, wherein the
biological sample is positioned between the X-ray source and the
flat panel detector, and wherein the X-ray source, the biological
sample, the flat panel detector, and the first camera are
collinear. In some embodiments, the method further comprises an
operation to convert the X-rays detected by the detection face of
the flat panel detector into a first X-ray image displayed on the
display face of the flat panel detector. In some embodiments, the
method further comprises an operation to record using the first
camera the first X-ray image displayed on the display face of the
flat panel detector. In some embodiments, the method further
comprises an operation to position the flat panel detector such
that the flat panel detector is not between the biological sample
and the camera. In some embodiments, the method further comprises
an operation to rotate the imaging stage by a predetermined amount
around at least of the first rotational axis and the second
rotational axis. In some embodiments, the method further comprises
an operation to position the flat panel detector between the
biological sample and the camera. In some embodiments, the method
further comprises an operation to irradiate the biological sample
on the rotatable imaging stage with X-rays using an X-ray source.
In some embodiments, the method further comprises an operation to
converting the X-rays detected through the transparent portion of
the rotatable imaging stage by the detection face of the flat panel
detector into a second X-ray image displayed on the display face of
the flat panel detector. In some embodiments, the method further
comprises an operation to record using the first camera the second
X-ray image displayed on the display face of the X-ray flat panel
detector.
[0080] FIG. 3 illustrates one embodiment as a descriptive example.
Shown is an apparatus 300 comprising a sample positioning module
301 and an optical imaging module 302. An X-ray flat panel detector
303 is positioned between a biological sample 304 and the optical
imaging module 302. A detection face 305 of the flat panel detector
303 is contacted by X-rays generated by an X-ray source 306 that
are not partially or completely blocked by regions of the
biological sample 304. The flat panel detector 303 converts the
X-rays detected by the detection face 305 into an X-ray image
displayed on the display face 307 of the flat panel detector. The
detection face 305 and the display face 307 are on opposite sides
of the flat panel detector 303. The optical imaging module 302 can
then be used to record the X-ray image displayed on the display
face 303. To record images of the biological sample 304 with the
optical imaging module 302 using imaging modalities other than
X-rays, the flat panel detector 307 is repositioned so that it is
not between the biological sample and the optical imaging module.
In this way, the optical imaging module 302 can record images
using, for example, one or both of a reflected visible light
channel or a fluorescence channel. The flat panel detector 303 can
be repeatedly moved from a first position enabling recording of
X-ray images with the optical imaging module 302 as described, to a
second position enabling recording of non-X-ray images with the
optical imaging module 302
[0081] In certain aspects, the method provides illuminating a
biological sample with visible light and capturing a plurality of
first 2-D images using visible light. The method further includes
illuminating the same or different biological sample with near
infrared light and using the camera to capture a plurality of
second 2-D images using infrared light. Preferably a single sample
is used so that both illumination techniques can be used
concurrently on a single sample without the visible light images
changing the appearance of the near infrared images or vice
versa.
[0082] In certain aspects, the plurality of 2-D first images are
taken at different angles of the imaging stage rotated through a
vertical axis. In certain other aspects, the plurality of 2-D first
images are taken at different angles of the imaging stage rotated
through a horizontal axis. In certain aspects, the plurality of 2-D
second images are taken at different angles of the imaging stage
rotated through a vertical axis. In certain aspects, the plurality
of 2-D second images are taken at different angles of the imaging
stage rotated through a horizontal axis.
[0083] FIG. 4 presents a flowchart of a process 400 for imaging a
biological sample with reflected visible light, fluorescence, and
ultrasound. In operation 401, a biological sample within an imaging
volume on a rotatable imaging stage is illuminated with visible
light, the rotatable imaging stage comprising a first rotational
axis, a second rotational axis, and a transparent portion, wherein
the second rotational axis is orthogonal to the first rotational
axis, and wherein the transparent portion is transparent to visible
light and near-infrared light. In operation 402, a first reflected
light image of visible light reflected by the biological sample is
recorded using a first camera. In operation 403, the biological
sample on the rotatable imaging stage is illuminated with
fluorescence excitation light using a fluorescence excitation light
source. In operation 404, a first fluorescence image of
fluorescence emission light emitted by the biological sample is
recorded using a second camera. In operation 405, energy pulses are
transmitted into the biological sample, wherein the energy pulses
are absorbed by the biological sample and converted into ultrasonic
emissions. In operation 406, the ultrasonic emissions are detected
using an ultrasonic transducer array. In operation 407, a first
ultrasound image constructed from the ultrasonic emissions detected
by the ultrasonic transducer array is recorded. In operation 408,
the imaging stage is rotated by a predetermined amount around at
least one of the first rotational axis and the second rotational
axis. In operation 409, a second reflected light image of visible
light reflected by the biological sample through the transparent
portion of the rotatable imaging stage is recorded. In operation
410, the biological sample is illuminated with fluorescence
excitation light. In operation 411, a second fluorescence image of
fluorescence emission light emitted by the biological sample
through the transparent portion of the rotatable imaging stage is
recorded. In operation 412, energy pulses are transmitted into the
biological sample, wherein the energy pulses are absorbed by the
biological sample and converted into ultrasonic emissions. In
operation 413, the ultrasonic emissions are detected using an
ultrasonic transducer array. In operation 414, a second ultrasound
image constructed from the ultrasonic emissions detected by the
ultrasonic transducer array is recorded.
[0084] In some embodiments, the method further comprises an
operation to construct a three-dimensional reflected light model
from the first and second reflected light images using a computer.
In some embodiments, the method further comprises an operation to
construct a three-dimensional fluorescence model from the first and
second fluorescence images using the computer. In some embodiments,
the method further comprises an operation to construct a
three-dimensional ultrasound model from the first and second
ultrasound images using a computer. In some embodiments, the method
further comprises an operation to render an image produced from the
reflected light model, the fluorescence model, and the ultrasound
model, wherein the reflected light model, the fluorescence model,
and the ultrasound model are identically registered in
three-dimensional space.
[0085] In some embodiments, the energy pulses are non-ionizing
laser pulses, and the ultrasound image is a photoacoustic image. In
some embodiments, the energy pulses are radio frequency pulses, and
the ultrasound image is a thermoacoustic image. In some
embodiments, the energy pulses are ultrasonic pulses.
[0086] FIG. 5 presents a flowchart of a process 500 for imaging a
biological sample with reflected visible light, fluorescence, and
optical coherence tomography. In operation 501, a biological sample
within an imaging volume on a rotatable imaging stage is
illuminated with visible light, the rotatable imaging stage
comprising a first rotational axis, a second rotational axis, and a
transparent portion, wherein the second rotational axis is
orthogonal to the first rotational axis, and wherein the
transparent portion is transparent to visible light and
near-infrared light. In operation 502, a first reflected light
image of visible light reflected by the biological sample is
recorded using a first camera. In operation 503, the biological
sample on the rotatable imaging stage is illuminated with
fluorescence excitation light using a fluorescence excitation light
source. In operation 504, a first fluorescence image of
fluorescence emission light emitted by the biological sample is
recorded using a second camera. In operation 505, the biological
sample on the rotatable imaging stage is illuminated with
near-infrared light. In operation 506, a first optical coherence
tomography image of near-infrared light reflected by the biological
sample is recorded using a third camera. In operation 507, the
imaging stage is rotated by a predetermined amount around at least
one of the first rotational axis and the second rotational axis. In
operation 508, a second reflected light image of visible light
reflected by the biological sample through the transparent portion
of the rotatable imaging stage is recorded. In operation 509, the
biological sample is illuminated with fluorescence excitation
light. In operation 510, a second fluorescence image of
fluorescence emission light emitted by the biological sample
through the transparent portion of the rotatable imaging stage is
recorded. In operation 511, the biological sample is illuminated
with near-infrared light. In operation 512, a second optical
coherence tomography image of near-infrared light reflected by the
biological sample is recorded.
[0087] In some embodiments, the method further comprises an
operation to construct a three-dimensional reflected light model
from the first and second reflected light images using a computer.
In some embodiments, the method further comprises an operation to
construct a three-dimensional fluorescence model from the first and
second fluorescence images using the computer. In some embodiments,
the method further comprises an operation to construct a
three-dimensional optical coherence tomography model from the first
and second optical coherence tomography images using a computer. In
some embodiments, the method further comprises an operation to
render an image produced from the reflected light model, the
fluorescence model, and the optical coherence tomography model,
wherein the reflected light model, the fluorescence model, and the
optical coherence tomography model are identically registered in
three-dimensional space.
[0088] In some embodiments, the method is used to image a
biological sample with visible light and fluorescence emissions.
Other imaging modalities that can be used with the method include
X-ray imaging to visualize tissue density and radiopaque tissue
inserts, photoacoustic imaging, optical coherence tomography,
ultrasound imaging, positron emission tomography, single-photon
emission computed tomography, Cherenkov luminescence imaging,
bioluminescence imaging, fluorescence lifetime imaging, and
spectroscopy.
[0089] FIG. 6 presents a flowchart of a process 600 for presenting
to an operator an image on a two-dimensional display. In operation
601, an image is displayed on a two-dimensional display, wherein
the image is a view of a subject from a viewpoint, wherein the
subject comprises a first rotational axis and a second rotational
axis, and wherein the second rotational axis is orthogonal to the
first rotational axis. The images from the viewpoint can be
constructed by overlaying, melding, or otherwise combining
reflected light, fluorescence, X-ray, ultrasound, and/or OCT images
taken as described above. In operation 602, the displayed image is
changed to a view of the subject from a viewpoint that is closer to
the subject in response to a zoom command by the operator. In
operation 603, the displayed image is changed to a view of the
subject from a viewpoint that is farther from the subject in
response to a pinch command by the operator. In operation 604, the
displayed image is changed to a view of the subject from a
viewpoint that is rotated around the first rotational axis in
response to a first rotational command by the operator. In
operation 605, the displayed image is changed to a view of the
subject from a viewpoint that is rotated around the second
rotational axis in response to a second rotational command by the
operator. In operation 606, information associated with at least a
portion of the displayed image is displayed in response to a
selection command by the operator
[0090] In some embodiments, the displayed image is produced from
two or more three-dimensional models. The models can be, for
example, any number of reflected light models, fluorescence models,
X-ray models, ultrasound models, and optical coherence tomography
models. Each model can be constructed from two or more images of
the subject. The models are typically identically registered in
three-dimensional space prior to producing the displayed image.
[0091] In some embodiments, one or more of the zoom, pinch,
rotational, or selection commands are entered using key presses,
control sticks, touch gestures, voice activation, or
accelerometers. In some embodiments, the commands are entered using
touch gestures. In some embodiments, the touch gestures are entered
using a touch pen.
[0092] In a surgical workflow, a surgeon who operates a surgery
only touches tools that are sterilized. In some surgical
procedures, a technologist or other staff member assists a surgeon
by helping to manipulate information presented on a display of any
instrument. However, actions taken by the staff may not accurately
or effectively accord with the verbal commands and requests from a
surgeon. As a result, there can be a benefit to enabling surgeons
to work with a display or instrument directly. Touching of
instruments such as a computer, keyboards, display panels, or a
cabinet imager may break the sterilization, though, and create
contamination problems. The use of a sterile touch pen to operate a
display or interface on a screen can therefore assist in
maintaining a sterile environment in an operating room.
[0093] FIG. 7 illustrates one embodiment as a descriptive example.
Shown is a touch pen 700 comprising a pen body 701 and a pen tip
702. The touch pen can also comprise a pen cover 703 that encloses
the pen body 701 and the pen tip 702.
[0094] The pen body 701 can be made of disposable and
pre-sterilized material intended for one-time or limited-time use.
The pen body 701 can be or made of sterilizable material intended
for repeated use with sterilization occurring prior to each use. In
some embodiments, one or both of the pen body 701 and the pen tip
702 comprise a metal. In some embodiments, the metal is stainless
steel. In some embodiments, the pen tip 702 is detachable from the
pen body 701. The pen tip 702 can be made of disposable and
pre-sterilized material intended for one-time or limited-time use.
The touch pen can be enclosed in a pen cover 703 that is made of
disposable and pre-sterilized material intended for one-time or
limited-time use. In some embodiments, the pen body 701 and pen tip
702 are not sterile, but the pen cover 703 is sterile. In some
embodiments, the touch pen can dispense ink from the pen tip. In
some embodiments, the touch pen does not dispense ink from the pen
tip.
[0095] In some embodiments, the touch pen has a touch tip at a
first end and an ink tip at a second end that is opposite to the
first end. The ink tip can be configured to dispense ink via, for
example and without limitation, a ballpoint tip, a rollerball tip,
a fountain tip, a felt tip, a small paint brush, or a spray
dispenser. A clinician can use the touch pen to mark on the
specimen directly. The mark can be used to indicate one or more
areas of interest and to refer to areas that have been identified
by the specimen imager and shown in the presentation display.
[0096] In some aspects, the pen dispenses visible ink. A clinician
can then use the touch pen, without switching to another pen, to
mark or put notes on a drape sheet, paper, towel, clothing article,
gauze, or other material present in the operating room or specimen
examination room. A clinician or other operator can also mark or
put notes on a container, cage, cartridge, or other supporting
material holding the excised specimen. Such notes are often kept
with the gross specimen for later reference. The use of the touch
pen for both operating the specimen imager and marking the sample
related to the imaging process can provide an advantage by
eliminating or reducing switching between a writing instrument and
a touch instrument. This can in turn help to avoid contaminations,
a particular concern for usage is in an operating room or other
substantially sterile environment.
[0097] In some aspects, the pen dispenses ink that includes a
fluorescent dye. The fluorescent ink can be visible or invisible.
The fluorescent dye can be any of those described above, and can,
for example, emit light in the NIR-I or NIR-II wavelength regions.
Such NIR ink can be used to mark areas of interest on the
biological sample or to provide annotations that can be visualized
with NIR imaging. In some embodiments, the NIR ink has a different
emission wavelength than that of fluorescent dyes used to label the
sample tissue. In these cases the NIR ink can be read with NIR
imaging, but will not interfere with readings of the staining of
the sample. In some embodiments, the NIR ink is used to apply the
staining to the sample. In these cases, the NIR ink can be selected
to, for example, target disease tissues or cancer cells. Such usage
can provide time and cost advantages by avoiding the systemic
application of label agents to large areas of the sample. Directed
application of NIR ink staining can also reduce the complexity of
background signal variations across the specimen, in particular
when the specimen comprises multiple tissue types. In some
embodiments, the touch pen is preloaded with one or more
fluorophores and targeting moieties.
[0098] The terms "about" and "approximately equal" are used herein
to modify a numerical value and indicate a defined range around
that value. If "X" is the value, "about X" or "approximately equal
to X" generally indicates a value from 0.90X to 1.10X. Any
reference to "about X" indicates at least the values X, 0.90X,
0.91X, 0.92X, 0.93X, 0.94X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X,
1.01X, 1.02X, 1.03X, 1.04X, 1.05X, 1.06X, 1.07X, 1.08X, 1.09X, and
1.10X. Thus, "about X" is intended to disclose, e.g., "0.98X." When
"about" is applied to the beginning of a numerical range, it
applies to both ends of the range. Thus, "from about 6 to 8.5" is
equivalent to "from about 6 to about 8.5." When "about" is applied
to the first value of a set of values, it applies to all values in
that set. Thus, "about 7, 9, or 11%" is equivalent to "about 7%,
about 9%, or about 11%."
[0099] Systems that incorporate the apparatus are also provided.
Systems can include, for example, power supplies, power regulators,
and other elements enabling the operation of the apparatus. It is
understood that the examples and embodiments described herein are
for illustrative purposes only and that various modifications or
changes in light thereof will be suggested to persons skilled in
the art and are to be included within the spirit and purview of
this application and scope of the appended claims. All
publications, patents, and patent applications, websites, and
databases cited herein are hereby incorporated by reference in
their entireties for all purposes.
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